KR100627643B1 - Pharmaceutical composition for treating or preventing type ii diabetes - Google Patents

Abstract

The present invention relates to an agent for treating or preventing type 2 diabetes, and more particularly, an agent for treating or preventing type 2 diabetes, comprising, as an active ingredient, mayis lignan or a pharmaceutically acceptable salt thereof represented by Formula (I) To a pharmaceutical composition.

[Formula I]

Macelignan, Insulin Resistance, Type 2 Diabetes  

Description

Pharmaceutical Composition for Treating or Preventing Type II Diabetes

1 is a process diagram for separating maceignan from Myristica fragrans .

Fig. 2 is the ¹³ C-NMR spectrum of the mace league of the present invention.

Figure 3 is a ¹ H-NMR spectrum of the mace league of the present invention.

Fig. 4 is the ¹ H- ¹ H COSY spectrum of the mayis lignan of the present invention.

Figure 5 is a ¹ H- ¹³ C HMBC spectrum of the maye lignan of the present invention.

Fig. 6 is the EI-Mass spectrum of the mace league of the present invention.

7 is a chemical structural formula of the mace league of the present invention.

8 is a result of reducing the blood glucose in the mace lignan of the present invention obesity / diabetes mouse model.

The present invention relates to a treatment or prophylactic agent of type 2 diabetes, and more specifically, mayrignan represented by the following formula (I) significantly reduced blood glucose in an obese / diabetic mouse model. Therefore, the mayis lignan of the present invention can be very useful for the treatment or prevention of type 2 diabetes.

[Formula I]

Diabetes can be roughly divided into type 1 and type 2 diabetes. Type 1 diabetes is an autoimmune disease caused by genetic susceptibility and viral infections resulting in an immunological response to the pancreas and selective destruction of β-cells. to be. Type 2 diabetes is the leading type of insulin resistance. The main etiologies are obesity, decreased insulin receptors, reduced tyrosine kinase activity, decreased muscle / adipose tissue type transporters in muscle and adipose tissue, and insulin receptor substrate-1 (IRS). -1) has been reported to be caused by complex causes such as intracellular deficiency. Type 2 diabetes occurs mainly in adults and accounts for 90% of all diabetes ( Drugs , 65: 433-445, 2005).

Myristica fragrans is a perennial plant grown in the tropics and its fruit, known as mace or nutmeg, has long been used as a spice. Macelignan is a typical lignan compound found in advance seutika Fragrance.. (Phytochemistry, 26: 1542-1543 , 1987) caspase (caspase) -3 activity enhancement action (Biol Pharm Bull of inducing apoptosis (apoptosis). , 27: 1305-1307, 2004), antimicrobial activity against oral microorganisms (Korean Patent Publication No. 10-2005-0035954), inhibitory effect on brain cell lipid peroxidation and free radical production ( Biochem. Biophys. Res. Commu. , 331 : 1264-1269, 2005). However, no studies have been reported on the anti-diabetic use of Mayslignan.

Therefore, the present inventors searched for various types of natural products to find an active substance that can treat or prevent insulin-resistant type 2 diabetes due to obesity, and mayis lignan isolated from mystica fragrance treats type 2 diabetes. And the present invention was completed by elucidating that it has an effect on prevention.

It is an object of the present invention to provide a novel use of mayis lignan isolated from Myristica fragrance extract as an agent for treating or preventing type 2 diabetes.

In accordance with the above object, the present invention provides a pharmaceutical composition for treating or preventing type 2 diabetes, comprising as an active ingredient mayis lignan represented by the following formula (I) or a pharmaceutically acceptable salt thereof.

[Formula I]

Hereinafter, the present invention will be described in more detail.

The present invention is characterized by providing a novel use of mayis lignan isolated from Myristica fragrance extract.

Mace lignan having antidiabetic activity of the present invention can be isolated and purified from myritica fragrance using a method of extracting and isolating conventional substances, preferably from nutmeg or aril. Can be purified. It can also be isolated and purified from another nutmeg plant, Myristica argentea Warb ( Nat. Prod. Lett ., 16: 1-7, 2002), and Machilus thunbergii ( Biol ). Pharm. Bull ., 27: 1305-1307, 2004), Leucas aspera , and the like, can also be isolated and purified ( Chem. Pharm. Bull ., 51: 595-598, 2003).

The mayis lignan of the present invention can be obtained by a conventional extraction and separation method. First, the dried mystica fragrance is placed in an extraction apparatus with 4 to 10 times of extraction solvent and left to extract for 2 days, and the concentrate is concentrated. Concentrate and dry to obtain an extract. In the present invention, as an extraction solvent for separation, water or an organic solvent of C1-C6 may be used. Preferably, purified water, methanol (methanol), ethanol, ethanol, propanol, isopropanol, butanol ( butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether Various solvents, such as (petroleum ether), can be used individually or in mixture. The extract obtained as described above may obtain a fraction according to polarity using silica gel column chromatography, and may separate the mayis lignan from the separated specific fraction through reverse phase column chromatography and high performance liquid chromatography (HPLC). However, the extraction and separation and purification method of the active ingredient is not necessarily limited to the above method.

Obesity / diabetic mice (ob / ob mouse) due to the lack of leptin (leptin) gene is not appetite is regulated to continue to excessive food intake. As a result, fat is excessively accumulated in the body, and at about 3 months after birth, it maintains around 50g, which is twice the weight of a typical mouse. It is also a model of typical type 2 diabetes with higher blood sugar levels compared to normal mice ( Exp. Clin. Endocrinol. Diabetes , 109: 307-319, 2001). These obese / diabetic mice are representative experimental animal models used to explore anti-obesity and anti-diabetic and therapeutic agents or to evaluate anti-obesity and anti-diabetic efficacy. The Mays lignan of the present invention suggested that it has a type 2 diabetes inhibitory effect by significantly reducing blood glucose in the obese / diabetic mouse model (see FIG. 8).

Mayes lignans according to the invention may be used in the form of salts, preferably pharmaceutically acceptable salts. The salt is preferably an acid addition salt formed with a pharmaceutically acceptable free acid. Organic acids and inorganic acids may be used as the free acid. The organic acid is not limited thereto, citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, metasulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, Glutamic acid and aspartic acid. In addition, the inorganic acid includes, but is not limited to, hydrochloric acid, bromic acid, sulfuric acid and phosphoric acid.

When administered clinically for the purpose of treating insulin resistant type 2 diabetes, the compositions of the present invention may be formulated into unit dosage forms suitable for oral or parenteral administration. When formulated in the form of a general medicine, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. which are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which solid preparations contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like. Are mixed to prepare. In addition to simple excipients, lubricants such as magnesium styrate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, ointments, creams. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used.

In addition, the composition of the present invention can be administered parenterally, parenteral administration is by subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method. To formulate into a parenteral formulation, mayis lignan is mixed in water with a stabilizer or buffer to prepare a solution or suspension, which is formulated in unit dosage forms of ampoules or vials. Dosage units may contain, for example, one, two, three or four times, or 1/2, 1/3 or 1/4 times the individual dosage. The individual dosage preferably contains an amount in which the effective drug is administered at one time, which usually corresponds to all, 1/2, 1/3 or 1/4 times the daily dose. Dosage levels for a particular patient may vary depending on the patient's weight, age, sex, health condition, diet, time of administration, method and excretion, drug combination and severity of the disease.

As a result of conducting oral toxicity test of the maye lignan of the present invention to rats, the 50% lethal dose (LD ₅₀ ) by oral toxicity test was found to be 2,000 mg / kg or more.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the following Examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto. In addition, all activity assays were repeated three or more times and the results were expressed as mean ± standard deviation.

Example 1 Isolation and Purification of Mayslignan from Myristica Fragrance

<1-1> Isolation and Purification of Mace Lignan

400 ml of 75% methanol was added to 100 g of dry ground nutmeg and left to stand at room temperature for two days. The extracted solution was filtered and concentrated in vacuo to prepare a nutmeg methanol extract (7 g), which was partitioned between ethyl acetate, butanol and water. The ethyl acetate fraction (4.2 g) was eluted with a solvent in which hexane and ethyl acetate were mixed at a ratio of 10: 1 (v / v) using silica gel column chromatography to obtain fraction III (1 g). Fraction III was column chromatographed with a solvent mixed with hexane and ethyl acetate at a ratio of 20: 1 (v / v) to give fraction III-B (0.52 g). Subsequently, fraction III-B was eluted with 80% methanol using Rp-18 column chromatography to obtain single substance fraction III-B-2 (0.5 g). Such a separation process chart is shown in FIG. 1 .

<1-2> Structure Analysis

In order to determine the structure of the isolated single material III-B-2, ¹ H-NMR spectrum and ¹³ C-NMR spectrum were measured at 600 MHz and 150 MHz (solvent: DMSO), respectively. The results are shown in FIGS . 2 and 3 , respectively. ¹³ C-NMR spectrum and the ^{1 H-NMR 1 H- 1 H} COSY spectrum and ¹ H- ¹³ to measure the correlation of the correlation of the ¹ H- ¹ H, based on the result of the spectrum related to the ¹ H- ¹³ C HMBC C The spectrum was measured. The results are shown in FIGS . 4 and 5 , respectively. ^{The results of 1} H-NMR, ¹³ C-NMR, ¹ H- ¹ H COZY, and ¹ H- ¹³ C HMBC were comprehensively analyzed and shown in Table 1 .

Position 13C-NMR 1H-NMR 1H-1H COSY 1H-13C HMBC One 135.4 2 109.2 6.72 brs C-7, C-6, C-4, C-3 3 147.3 4 145.1 5 107.9 6.79 d (7.8) 6.61 C-6, C-4, C-3, C-1 6 121.7 6.61 dd (7.8) 6.79 C-7, C-5, C-4, C-2, C-1 7 38.2 2.23 dd (13.2, 9.3) 2.66 dd (13.2, 4.8) 1.64, 2.66 1.64, 2.23 C-8, C-6, C-2, C-1 C-9, C-8, C-6, C-2, C-1 8 38.7 1.64 brs 0.75, 2.23, 2.66 C-7 9 16.0 0.75 d (6.3) 1.64 C-8, C-7 One' 132.4 2' 112.9 6.66 brs C-7 ', C-6', C-4 ', C-3' 3 ' 147.1 4' 144.4 5 ' 115.2 6.66 d (7.9) 6.53 C-6 ', C-4', C-3 ', C-1' 6 ' 121.0 6.53 d (7.9, 1.1) 6.66 C-7 ', C-5', C-4 ', C-2', C-1 ' 7 ' 38.0 2.17 dd (13.2, 9.3) 2.66 dd (13.2, 4.8) 1.64, 2.66 1.64, 2.17 C-8 ', C-6', C-2 ', C-1' C-9 ', C-8', C-6 ', C-2', C-1 ' 8' 38.7 1.64 brs 0.75, 2.17, 2.66 C-7 ' 9 ' 16.1 0.75 d (6.3) 1.64 C-8 ', C-7' OMe 55.5 3.72 (s) O-CH2-O 100.6 5.95 d (4.8) C-3, C-4

<1-3> mass spectrometry

The results of EI / MS measured for mass spectrometry of the separated single substance are shown in FIG. 6 . The compound had a molecular weight of 328 with [M] ⁺ being observed at m / z 328 in EI / MS, and the molecular formula was C ₂₀ H ₂₄ O ₄ .

<1-4> Non-photon measurement

20 mg of the separated single substance III-B-2 was dissolved in 2 ml of chloroform (CHCl ₃ ), and then the specific light intensity ([α] _D ) value was measured by a polarimeter (Automatic Polarimeter, APIII-589, Rodulph, NJ, USA). [α] _D = +4.0 (CHCl ₃ , c = 1.0).

Results of ¹ H-NMR, ¹³ C-NMR, ¹ H- ¹ H COZY, ¹ H- ¹³ C HMBC, EI / MS and [α] _D and previously published research reports ( Phytochemistry , 26: 1542-) 1543, 1987) was identified and compared, it was confirmed that the isolated single material is macelignan ( Fig. 7 ).

Example 2 Antidiabetic Effect of Mayslignan in Obese / Diabetes Mouse Model

  Obesity / diabetic mice used as a diabetic model animals are inadequately controlled due to a lack of leptin genes, resulting in excessive food intake. As a result, fat is excessively accumulated in the body and becomes a model of the typical type 2 diabetes which has a high blood sugar level compared with a normal mouse. Therefore, four 10-week-old obese / diabetic mice were used in each experimental group to investigate the prevention and treatment of diabetes. Each dose was administered at a constant time for 10 days, and the control group received only the same amount of corn oil. At 10 days after the start of administration, blood glucose levels of the test and control groups were measured and analyzed.

As shown in FIG. 8 , blood glucose was 100% (420.63 ± 10.66 mg / dl) in the control group and 49.13% (206.65 ± 19.95 mg in normal mice). compared to the maintenance of hyperglycemia compared to that maintained in the hyperglycemia group, the group treated with Mayslignan showed a significant decrease in blood sugar levels (*, p <0.01) of 71.8% (298.8 ± 30.08 mg / dl) to prevent diabetes. And the treatment effect was observed, and mayslignan showed a much higher hypoglycemic effect than troglitazone, which is currently used as a diabetes treatment.

Example 3 Antidiabetic Effect of Ethanol Extract in Obese / Diabetes Mouse Model

400 ml of ethanol was added to 100 g of dry ground nutmeg and left to stand at room temperature for two days. The extracted solution was filtered and concentrated in vacuo to prepare nutmeg ethanol extract (6 g).

As a result of assaying the anti-diabetic effect in the obese / diabetic mouse model in the same manner as in Example 5 with respect to the prepared ethanol extract, blood sugar was reduced by 14.5% compared to the control.

Example 4 Antidiabetic Effect of Hexane Extract in Obese / Diabetes Mouse Model

400 ml of hexane was added to 100 g of dry ground nutmeg, and the mixture was left at room temperature for two days. The extracted solution was filtered and concentrated in vacuo to prepare a nutmeg hexane extract (14 g).

As a result of assaying the anti-diabetic effect in the obese / diabetic mouse model in the same manner as in Example 5 with respect to the prepared hexane extract, blood sugar was reduced by 12.7% compared to the control.

<Manufacture example 1>

Preparation of a medicament containing the pharmaceutical composition for treating or preventing diabetes mellitus according to the present invention

<1-1> Preparation of Tablet

25 mg of the lignan compound or myristica fragrance extract of the present invention, 26 mg of lactose for excipients and 3.5 mg of avicel (microcrystalline cellulose), 1.5 mg of sodium starch glyconate as a disintegration aid, and L-HPC for direct-acting agent (low) -hydroxypropylcellulose) was added to a U-type mixer with 8 mg and mixed for 20 minutes. After mixing was completed, 1 mg of magnesium stearate was further added as a lubricant and mixed for 3 minutes. Tablets were prepared by tableting and film coating after a quantitative test and a humidity test.

<1-2> Preparation of Syrup

A syrup containing maye lignan of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient of 2% (W / V) was prepared by the following method: 2 g of acid addition salt of maye lignan of the present invention, 0.8 g of saccharin, and 25.4 g of sugar was dissolved in 80 g of warm water. After cooling the solution, 8.0 g of glycerin, 0.04 g of flavor, 4.0 g of ethanol, 0.4 g of sorbic acid, and an appropriate amount of distilled water were mixed thereto. Water was added to the mixture to make 100 mL.

<1-3> Preparation of Capsule

A capsule was prepared by mixing 50 mg of lignan compound or myristica fragrance extract of the present invention, 50 mg of lactose, 46.5 mg of starch, 1 mg of talc, and an appropriate amount of magnesium stearate and filling it into a hard gelatin capsule.

<1-4> Preparation of Injection Solution

Injectable solutions containing 10 mg of the active ingredient were prepared by the following method: 100 ml were prepared by dissolving 1 g of mayis lignan hydrochloride, 0.6 g of sodium chloride, and 0.1 g of ascorbic acid in distilled water. The solution was bottled and sterilized by heating at 20 ° C. for 30 minutes.

As described above, the Mays lignan and Myristica fragrans extracts of the present invention have a significant effect on reducing blood sugar in obese / diabetic animal models, and thus may be usefully used as preventive and therapeutic agents for insulin resistant type 2 diabetes. Can be.

Claims (2)

A pharmaceutical composition for treating or preventing type 2 diabetes, comprising mayis lignan represented by the following formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient: [Formula I]

Pharmaceutical composition for the treatment or prevention of type 2 diabetes, characterized by extracting Myristica fragrans as an active ingredient

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